title: The presence of outer arm fucose residues on the N-glycans of tissue inhibitor of metalloproteinases-1 reduces its activity.
abstract: Tissue inhibitor of metalloproteinases-1 (TIMP-1) inhibits matrix metalloproteinases (MMPs) by binding at a 1:1 stoichiometry. Here we have shown the involvement of N-glycosylation in the MMP inhibitory ability of TIMP-1. TIMP-1, purified from HEK 293 cells overexpressing TIMP-1 (293 TIMP-1), showed less binding and inhibitory abilities to MMPs than TIMP-1 purified from fibroblasts or SF9 insect cells infected with TIMP-1 baculovirus. Following deglycosylation of TIMP-1, all forms of TIMP-1 showed similar levels of MMP binding and inhibition, suggesting that glycosylation is involved in the regulation of these TIMP-1 activities. Analysis of the N-glycan structures showed that SF9 TIMP-1 has the simplest N-glycan structures, followed by fibroblast TIMP-1 and 293 TIMP-1, in order of increasing complexity in their N-glycan structures. Further analyses showed that cleavage of outer arm fucose residues from the N-glycans of 293 TIMP-1 or knockdown of both FUT4 and FUT7 (which encode for fucosyltransferases that add outer arm fucose residues to N-glycans) enhanced the MMP-binding and catalytic abilities of 293 TIMP-1, bringing them up to the levels of the other TIMP-1. These results demonstrate that the ability of TIMP-1 to inhibit MMPs is at least in part regulated by outer arm fucosylation of its N-glycans.
authors.affiliation: Department of Biochemistry, College of Science and Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Han Ie
authors.initials: HIe
authors.lastname: Kim
authors.forename: Radka
authors.initials: R
authors.lastname: Saldova
authors.forename: Jun Hyoung
authors.initials: JH
authors.lastname: Park
authors.forename: Young Hun
authors.initials: YH
authors.lastname: Lee
authors.forename: David J
authors.initials: DJ
authors.lastname: Harvey
authors.forename: Mark R
authors.initials: MR
authors.lastname: Wormald
authors.forename: Kieran
authors.initials: K
authors.lastname: Wynne
authors.forename: Giuliano
authors.initials: G
authors.lastname: Elia
authors.forename: Hwa-Jung
authors.initials: HJ
authors.lastname: Kim
authors.forename: Pauline M
authors.initials: PM
authors.lastname: Rudd
authors.forename: Seung-Taek
authors.initials: ST
authors.lastname: Lee
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001665:Binding Sites; D002240:Carbohydrate Sequence; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005643:Fucose / Q000737:chemistry* / Q000378:metabolism; D005647:Fucosyltransferases / Q000172:deficiency / Q000235:genetics; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D016256:Lewis X Antigen / Q000235:genetics; D020781:Matrix Metalloproteinase 1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D061987:Sf9 Cells; D015398:Signal Transduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D018411:Spodoptera; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D016256:Lewis X Antigen; D011134:Polysaccharides; D011994:Recombinant Proteins; C560822:TIMP1 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D005643:Fucose; C066693:FUT4 protein, human; C502597:FUT7 protein, human; D005647:Fucosyltransferases; D020781:Matrix Metalloproteinase 1